OBJECTIVE:To isolate and purify three principal degraded impurities of mitiglinide calcium (impurity A,B,C) and identify their structures,establish HPLC method for content determination of impurity A,B,C. METHODS:Mitiglinide calci-um was used as raw material and reacted with acid;3 impurities were then separated by HPLC and their structures were elucidated by IR,MS,1H NMR,13C NMR,LC-ESI-MS and ORD. 3 impurities of 3 batches of mitiglinide calcium were determined,and the determination was performed on Agilent Extend-C18 column with mobile phase consisted of 0.01 mol/L sodium acetate solution-ace-tonitrile-triethylamine(60:40:0.1,pH=3.0)at the flow rate of 1.0 ml/min. The detection wavelength was set at 210 nm and sam-ple size was 20 μl. The response tests of 3 impurities and mitiglinide calcium were conducted. RESULTS:After treated with acid, impurity A,B,C had been obtained,and their purity were 99.05%,98.87%,99.98%,respectively after isolation and purifica-tion;after identifying the structure, 3 impurities were S-2-bezylsuccinic acid, S-2-bezylsuccinic acid-4-methyl ester, methyl (2S)-2-benzyl-3-(cis-hexahydroisoindolin-2-ylcarbonyl) propionate;methodological study of content determination of impurities were all up to the requirement. The linear range of impurity A,B,C were 0.387 5-3.875,0.395-3.95 and 0.392 5-3.925 μg/ml(all r were 1.000 0). The response value of impurity A,B,C and mitiglinide calcium were 2.316 1,2.636 1,2.617 8 and 2.620 4,re-spectively. CONCLUSIONS:The structures of 3 principal degraded impurities of mitiglinide calcium have been identified and con-firmed;the content of them can be determined by HPLC main component self-comparison method.

Aim To study the inhibitory effects of ex-tractive pericarpium trichosanthes ( EPT) on high glu-cose-induced apoptosis in human umbilical vein endo-thelial cells ( HUVECs ) and its underlining mecha-nisms. Methods HUVECs were cultured. Effects of EPT at different concentrations on the high-glucose-in-duced apoptosis in HUVECs were observed. The cell viability of HUVECs was determined by MTT colori-metric method. Cell apoptosis was identified by Ho-echst staining. The intracellular activity of Caspase-3 was detected with colorimetry. Protein expression and p65 nuclear translocation of NF-κB were detected by Western blot and immunofluorescence staining. Re-sults Treated with 30 mmol · L-1 glucose for 48 hours had a significantly decrease on cell viability com-pared with control, and the apoptotic rate and Caspase-3 activity were increased markedly, the protein expres-sion of NF-κB was upregulated and p65 nuclear trans-location in HUVECs increased; Pre-incubation with EPT(12. 5,25,50 mg·L-1 ) for 1 hour enhanced the cell viability, and decreased the apoptotic rate and Caspase-3 activity and downregulated the expression of NF-κB in a dose-dependent manner. Moreover, EPT could inhibit p65 translocation. Conclusion EPT can protect HUVECs against the apoptosis induced by high glucose in vitro,and its mechanism may be related with downregulation of NF-κB expression and inhibition of the intracellular activity of Caspase-3 .

Objective To observe the effect of acupuncture penetrated from Tiaokou (ST38) to Chenshan (BL57) combined with exer-cise on affected limbs of shoulder-hand syndrome (SHS) at stage I after stroke. Methods 40 inpatients with SHS at stage I after stroke from January, 2013 to December, 2014 were randomly divided into treatment group (n=20) and control group (n=20). The treatment group accept-ed acupuncture from Tiaokou to Chenshan on unaffected sides and exercise of affected upper limbs, and the control group accetpted routine acupuncture on Triple Shoulder Acupoints on affected sides. They were assessed with Pain Rating Index (PRI), Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) before and 4 weeks after intervention. Results The incidence of improvement was 90% in the treatment group, and 80%in the control group (χ2=0.784, P=0.376). The scores of PRI decreased (t>5.37, P<0.001), and the score of FMA and MBI increased (t>4.66, P<0.001) significantly after intervention. The score of FMA and MBI improved more in the treatment group than in the control group (t>2.25, P<0.05). Conclusion Acupuncture from Tiaokou to Chenshan combined with exercise is more effective on SHS at stage I after stroke than conventional Triple Shoulder Acupoints.

Objective To explore the expression and role of LAT 1 in mouse uterus on early placenta formation on day 8 of pregnancy (D8).Methods One hundred and twenty 6-8-week old SPF female Kunming mice were used in this study.Immunohistochemistry was applied to determine the localization of LAT 1 protein in the mouse uterus on D8 of preg-nancy.The ectoplacental cones (EPCs) were dissected out from D8.5 uterus, and then cultured in vitro with different con-centrations of BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, specific antagonist of LAT1) and L-leucine ( substrate of LAT1) to determine the role of LAT1 during the EPC attachment and outgrowth .Results LAT1 protein was highly expressed in secondary decidual zone and also positively expressed in the mouse uterus on D 8.As a specific antago-nist of LAT1, BCH significantly suppressed the ectoplacental cone outgrowth , whereas L-leucine showed no significant effect on it.Conclusions LAT1 is expressed in the mouse uterus during early placenta formation and promotes ectoplacen -tal cone outgrowth , suggesting that LAT1 may promote the trophoblast invasion into maternal decidual tissue , and partici-pates in the early formation of placenta .

Objective:To design and construct the replication-deficient recombinant adenovirus Ad-siCTGF which can silence the expression of connective tissue growth factor (CTGF) by RNA interference and verified its function. Methods:A specific sequence, which was verified to be able to silence CTGF gene with high efficiency, was cloned into pSES-HUS vector to produce the shuttle plasmid pSES-siCTGF. The plasmid after Pme Ⅰ linearization was cotransduced with pAdEasy into BJ5183 E.coli strains to construct recombinant vector Ad-siCTGF. After linearization treatment with Pac Ⅰ enzyme digestion Ad-siCTGF was transfected into HEK293 cells via liposome mediation. The recombinant adenovirus was packaged. The titer of the Ad-siCTGF was increased after three times of cross-infection. 4T1 cells were infected with the adenovirus. The silencing efficiency was tested by real-time fluorescence quantitative (RFQ)-PCR and Western blotting.Results:Pac Ⅰ enzyme digestion electrophoresis indentified that recombinant adenovirus was successfully constructed. The titer of the recombinant adenovirus Ad-siCTGF was 2.6×10~(10) pfu/mL after amplification and purification. The CTGF mRNA and protein expression in 4T1 cells were decreased by 36.27% and 31.56%, respectively, compared with the control groups.Conclusion:The recombinant adenovirus which can silence the expression of CTGF was successfully constructed. It laid a good foundation for further investigation of the action mechanism of CTGF in tumor cells.

Pulmonary vein isolation (PVI) is a new ,effective and radical method to cure atrial fibrillation .Within a period after radiofrequency ablation (RFA) ,coagulation system is activated in patients ,then thrombus incident such as cerebral embolism may happen .The present article made a review on its mechanism .

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .

Objective:To observe the adverse reactions of killer cytokine-induced (CIK) in the treatment of malignant tumor and to analyze the possible mechanism ,and to develop the targeted prevention and treatment measures .Methods: The clinical data, including various adverse reactions , laboratory tests and the corresponding preventive measures against adverse reactions .In 1 240 patients with malignant tumor after treated with CIK cells from May 2013 to September 2015 were retrospectively analyzed .Results:The main adverse reactions after the first infusion of CIK cells were weak (10%),fever(7.25%),shiver (4%),arthralgia (3%),systemic in flammatory response syndrome reaction ( 3%) , digestive tract discomfort ( 0.96%) , acute allergic reaction ( 0.08%) , rash (0.08%),angina pectoris (0.08%),tumor lysis syndrome(0%),infection(0%).With the increase of the treatment ,the incidence of adverse reactions increased and the fever was the main performance ,after the fourth course into the platform .The combination of blood pressure increased or decreased and severe allergic reaction and systemic inflammatory response syndrome was needed to be treated .The CIK cells were pretreated before treatment could reduce the incidence of these reactions .Conclusion:CIK cells therapy is a safe and effective adoptive immunotherapy for malignant tumor and its adverse reactions can be treated expectantly , but rare adverse reactions may have potential risks .

To identify the structure of three related substances in potassium sodium dehydroandrographolide succinate (PSDS), an HPLC preparation method was used to separate the impurities. These main impurities were identified using LC-ESI/TOFMS, LC-ESI/MSn, NMR, UV and IR. One of the main impurities was a hydrolyzed and oxidized product of PSDS, which has not been reported previouely. The other two impurities were hydrolyzed products of PSDS after losing different succinic acids. The results indicate that PSDS can be easily hydrolyzed and oxidized. It should be stored at cool and dry places.

With the development of civil military integration,military scientific research institu tes are facing the challenge of constructing a new mode of translating scientific and technological achievements into practice and enhancing translational efficiency.This paper began with the evolution of translation mode in military institutes and discussed the flaws and insufficiency of current mode,then a triple helix translation mode,which encompass government,industry and research,was introduced and fully explained for future reference.